Background: Staphylococcus pseudintermedius (SP) is the major pathogenic species of dogs involved in a wide variety of skin and soft tissue infections. The accessory gene regulator (agr) locus of Staphylococcus aureus has been extensively studied, and it influences the expression of many virulence genes. It encodes a two-component signal transduction system that leads to down-regulation of surface proteins and up-regulation of secreted proteins during in vitro growth of S. aureus. The objective of this study was to detect and sequence analyzing the AgrA, B, and D of SP isolated from canine skin infections.

Materials and Methods: In this study, we have isolated and identified SP from canine pyoderma and otitis cases by polymerase chain reaction (PCR) and confirmed by PCR-restriction fragment length polymorphism. Primers for SP agrA and agrBD genes were designed using online primer designing software and BLAST searched for its specificity. Amplification of the agr genes was carried out for 53 isolates of SP by PCR and sequencing of agrA, B, and D were carried out for five isolates and analyzed using DNAstar and Mega5.2 software.

Results: A total of 53 (59%) SP isolates were obtained from 90 samples. 15 isolates (28%) were confirmed to be methicillinresistant SP (MRSP) with the detection of the mecA gene. Accessory gene regulator A, B, and D genes were detected in all the SP isolates. Complete nucleotide sequences of the above three genes for five isolates were submitted to GenBank, and their accession numbers are from KJ133557 to KJ133571. AgrA amino acid sequence analysis showed that it is mainly made of alpha-helices and is hydrophilic in nature. AgrB is a transmembrane protein, and AgrD encodes the precursor of the autoinducing peptide (AIP). Sequencing of the agrD gene revealed that the 5 canine SP strains tested could be divided into three Agr specificity groups (RIPTSTGFF, KIPTSTGFF, and RIPISTGFF) based on the putative AIP produced by each strain. The AIP of SP contains serine and produce lactone ring structured AIP.

Conclusion: Presence of AgrA, B, and D in all SP isolates implies the importance of this regulatory system in the virulence genes expression of the SP bacteria. SP isolates can be typed based on the AgrD auto-inducible protein sequences as it is being carried out for typing of S. aureus isolates. However, further studies are required to elucidate the mechanism of controlling of virulence genes by agrgene locus in the pathogenesis of soft tissue infection by SP.

Aim: The present study was aimed to investigate the relationship of udder shape, teat-end shape, teat length, and teat diameter with intra-mammary infection in Jersey crossbred cows under hot-humid climate.

Materials and Methods: A total of 24 lactating Jersey crossbred cows were evaluated for udder shape (pendulous/regular) and teat-end shape (flat/inverted/pointed) by visual examination, while teat length and teat diameter were measured using vernier caliper. Monthly milk sampling was done for 4 months of duration. Few quarters were found as blind or nonfunctional and so, a total of 366 quarter wise milk samples were collected at the monthly interval and subjected to somatic cell count (SCC) microscopically. The data on SCC were transformed into log scale and analyzed.

Results: There was a significant (p<0.01) effect of udder shape and teat-end shape on SCC level. The mean SCC level for pendulous udder was significantly (p<0.05) higher as compared to the regular shaped udder. Similarly, significantly (p<0.05) a higher level of mean SCC was found in flat teat-end shape. A significant (p<0.01) correlation was found between SCC and teat length as well as teat diameter.

Conclusion: In conclusion, pendulous udder, flat and inverted teat-end, very long and thick teat were more susceptible to intra-mammary infection in Jersey crossbred cows and these traits must be considered accordingly while selecting dairy animals for future milk production.

Aim: One of the major biochemical aspects of thermoregulation is equilibrium of ion gradient across biological membranes. Na+/K+-ATPase, a member of P type-ATPase family, is a major contributor to the mechanism that actively controls crossmembrane ion gradient. Thus, we examined ATP1A1 gene that encodes alpha-1 chain of Na+/K+-ATPase, for genetic polymorphisms.

Materials and Methods: A total of 100 Vrindavani (composite cross strain of Hariana x Holstein-Friesian/Brown Swiss/Jersey) and 64 Tharparkar (indigenous) cattle were screened for genetic polymorphism in ATP1A1 gene, using polymerase chain reaction single-strand conformation polymorphism and DNA sequencing. For association studies, rectal temperature (RT) and respiration rate (RR) of all animals were recorded twice daily for 3 seasons.

Results: A SNP (C2789A) was identified in exon 17 of ATP1A1 gene. Three genotypes namely CC, CA, and AA were observed in both, Vrindavani and Tharparkar cattle. The gene frequencies in Tharparkar and Vrindavani for allele A were 0.51 and 0.48, and for allele C were 0.49 and 0.52, respectively, which remained at intermediate range. Association study of genotypes with RT and RR in both cattle population revealed that the animals with genotype CC exhibited significantly lower RT and higher heat tolerance coefficient than CA and AA genotypes.

Conclusion: Differential thermoregulation between different genotypes of ATP1A1 gene indicate that the ATP1A1 gene could be potentially contributing to thermotolerance in both, Tharparkar, an indigenous breed and Vrindavani, a composite crossbred cattle.

Nano minerals are widely used in diversified sectors including agriculture, animal, and food systems. Hence, their multiple uses provoke the production of nanomaterials at the laboratory level, which can be achieved through physical, chemical or biological methods. Every method is having its own merits and demerits. But keeping all in mind, chemical methods are more beneficial, as uniform nano-sized particles can be produced, but the use of corrosive chemicals is the main demerits. When it comes to environmental issues, biological methods are better as these are free from corrosive chemicals, but maintaining the culture media is the disadvantage. For animal feeding, chemical methods are mostly followed to produce nano minerals as it is cheap and less time consuming. These nano minerals also showed their significant effects even at lower doses of recommendations than the conventional mineral sources. These nano minerals have significant growth promoting, immuno-modulatory, antibacterial effects than the conventional counterparts. They also alter the rumen fermentation pattern on supplementation in the animal feeds. Apart from these, nano minerals are reported to enhance the reproduction in the livestock and poultry.

Aim: The objective of the study was to evaluate the effect of deep litter housing and fermented feed on carcass characteristics and meat quality of crossbred Hampshire pigs.

Materials and Methods: Forty-eight weaned crossbred Hampshire piglets of 2 months age (24 males and 24 females) were selected for the experiment. The piglets were randomly assigned into 4 homogenous experimental groups with 6 males and 6 females each: E1; reared on a conventional housing and fed with a fermented diet, E2; reared on a deep litter housing system and fed with a fermented diet, E3; reared on a deep litter housing system and fed with a conventional diet and C; reared on a conventional housing system and fed with a conventional diet. The study was continued up to 32 weeks of age and at the end of this period, 6 animals (3 males and 3 females) from each experimental group were slaughtered to evaluate carcass traits and meat quality characteristics.

Results: Pre-slaughter weight, hot carcass weight, and dressing percentage were significantly (p<0.01) affected by feeding fermented diet and deep litter housing while carcass traits, i.e., carcass length, backfat thickness, and loin eye area were not affected. The edible offal; liver and heart weight (p<0.05) differed significantly while kidney weight showed no difference. The inedible offal; head weight (p<0.01) and lung weight revealed a significant difference (p<0.05) while spleen and stomach weight showed no difference among the experimental groups. The wholesale cuts and meat: bone ratio of pigs also differed significantly among the groups. Morphometry of small and large intestine also showed a significant difference. Chemical composition of pork viz., moisture and total ash content was influenced by the treatment, while crude protein and ether extract content were not affected. Mineral composition of pork also showed no significant difference. Color characteristics of Longissimus dorsi muscle showed a significant difference in L* and a* value while parameter b* was not affected. The tenderness of meat showed significant difference among the groups (p<0.01).

10. Molecular identification and genetic diversity of open reading frame 7 field isolated porcine reproductive and respiratory syndrome in North Sumatera, Indonesia, in the period of 2008-2014 - Faisal Faisal, Rini Widayanti, Aris Haryanto and Charles Rangga Tabu

Veterinary World, 8(7): 875-880

doi: 10.14202/vetworld.2015.875-880

Abstract

Aim: Molecular identification and genetic diversity of open reading frame 7 (ORF7) of field isolated porcine reproductive and respiratory syndrome virus (PRRSV) in North Sumatera, Indonesia, in the period of 2008-2014.

Materials and Methods: A total of 47 PRRSV samples were collected from the death case of pigs. The samples were collected from different districts in the period of 2008-2014 from North Sumatera province. Two pairs of primer were designed to amplify ORF7 of Type 1 and 2 PRRSV based on the sequence of reference viruses VR2332 and Lelystad. Viral RNAs were extracted from samples using PureLink™ micro-to-Midi total RNA purification system (Invitrogen). To amplify the ORF7 of PRRSV, the synthesis cDNA and DNA amplification were performed by reverse transcription polymerase chain reaction (RT-PCR) and nested PCR method. Then the DNA sequencing of PCR products and phylogenetic analysis were accomplished by molecular evolutionary genetics analysis version 6.0 software program.

Results: RT-PCR and nested PCR used in this study had successfully detected of 18 samples positive PRRS virus with the amplification products at 703bp and 508bp, respectively. Sequencing of the ORF7 shows that 18 PRRS viruses isolated from North Sumatera belonged to North American (NA). JXA1 Like and classic NA type viruses. Several mutations were detected, particularly in the area of nuclear localization signal (NLS1) and in NLS2. In the local viruses, which were related closed to JXA1 virus; there are two differences in amino acids in position 12 and 43 of ORF7. Our tested viruses showed that the amino acid positions 12 and 43 are Asparagine and Arginine, while the reference virus (VR2332, Lelystad, and JXA1) occupied both by Lysine. Based on differences in two amino acids at position 12 and 43 showed that viruses from North Sumatera has its own uniqueness and related closed to highly pathogenic PRRS (HP-PRRS) virus (JXA1).

Conclusion: The results demonstrated that North Sumatera type PRRS virus has caused PRRS outbreaks in pig in North Sumatera between 2008 and 2014. The JAX1 like viruses had unique amino acid residue in position 12 and 43 of asparagine and lysine, and these were genetic determinants of North Sumatera viruses compared to other PRRS viruses.

Aim: This study was undertaken to derive the prevalence rate of Fasciolosis in buffaloes by a collection of fecal and liver samples from Anand and Ahmedabad districts’ local slaughter houses.

Materials and Methods: Fecal and liver samples were collected during ante- and post-mortem examination, respectively, and brought to the department laboratory preserved in 10% formalin for further processing. Fecal samples were processed with qualitative examinationviz.; sedimentation technique for identification of the ova. Liver samples were also examined for the presence of gross parasites.

Results: The highest prevalence rate was observed in the month of December (25.97% fecal and 33.33% liver samples) and lowest in the month of May (10.71% fecal and 11.76% liver samples) at Anand district. In the area of Ahmedabad district, the highest prevalence rate was recorded in the month of October and February (26.98%) and lowest in the month of May (10.34%) for the fecal and highest prevalence was observed in the month of February (26.98%) and lowest in May (11.11%) for the liver samples.

Conclusion: It can be concluded that the heavy infection is present in Anand and Ahmedabad districts, especially in the month of winter followed by monsoon and the least in summer.

Aim: This study was conducted to determine the seroprevalence and distribution of foot and mouth disease virus (FMDV) infection in pigs from Zuru, Kebbi State, Nigeria.

Materials and Methods: Cross-sectional serological surveys were conducted between May and December 2013 using the immunochromatography assay technique. Structured questionnaires were administered to households identified at pig rearing areas to obtain the population structures and some information on managemental practices.

Results: A total number of 849 pigs were enumerated at 37 pigs rearing households. Tudun wada had the largest concentration of pigs (237 pigs), while Dabai has the least (38 pigs). A total of 250 blood samples were collected, of which 45 (18%) were positive; Zango has the highest seroprevalence (1.6%), while Dabai recorded the least (0.4%). Based on sex and age, the infection was higher in female (10.4%) and young pigs (11.6%) than male (7.6%) and adult pigs (6.4%), respectively. There was no significant (p>0.05) association between infection and pig rearing areas, sex, and age. Furthermore, none of the 250 pigs examined for classical FMDV lesions was positive.

Conclusion: The results of this study showed that FMD is an important disease of pigs in the study areas. This result justifies the need for more attention and subsequent molecular study to identify the circulating FMDV in the area, which will help in the implementation of effective control measures.

Aim: This study assessed the impact of four water sources used as drinking water in Egypt for broiler chickens on its performance, carcass characteristic, hematological, and immunological responses.

Materials and Methods: A total of 204 unsexed 1-day old Indian River broiler chickens were used in this study. They were randomly allocated into four treatment groups of 51 birds in each, with three replicates, 17 birds per replicate. Groups were classified according to water source they had been received into (T1) received farm tap water; (T2) received filtered tap water (T3) received farm stored water at rooftop tanks, (T4) received underground (well) water.

Conclusion: Base on the results of this study, it is concluded that different water sources could safely be used as drinking water for poultry; as long as it is present within the acceptable range of drinking water quality for chickens. Suggesting the benefits of treatment of water sources on improving chickens’ health and welfare. Draw attention to the importance of maintaining the hygienic quality of stored water.

Aim: To estimate existing within-breed genetic variability in Marwari goats under field conditions and the generated data that can be used to determine genetic relationships with other breed of goats.

Materials and Methods: A total of 146 blood samples of goats of Marwari breed were randomly collected from genetically unrelated animals from different villages of Bikaner Districts of Rajasthan, India. Genomic DNA was extracted from whole blood using proteinase K-digestion followed by standard phenol–chloroform extraction procedure at room temperature and confirmed through horizontal electrophoresis on 0.8% agarose gel containing ethidium bromide. Fifteen caprine microsatellite markers were used to estimate genetic variability among the goats of Marwari breed in terms of allelic and genotype frequencies, heterozygosities and polymorphism information content (PIC) value.

Results: A total of 74 alleles were contributed by Marwari goat across all 15 microsatellite loci. The number of alleles per locus varied from two (ILSTS-087) to 9 (ILSTS-058) alleles, with a mean of 4.93 whereas the effective number of allele varied from 1.35 (ILSTS-005) to 3.129 (ILSTS011) with a mean of 2.36. The effective number of allele is lesser than observed number at all the loci. Allelic sizes ranged from 125 bp (ILSTS-028 and ILSTS-033) to 650 bp (ILSTS-011 and ILSTS-019). The expected heterozygosity ranged from 0.240 (locus ILSTS-005) to 0.681 (locus ILSTS-011), with an average value of 0.544. The observed heterozygosity (Ho) ranged from 0.1428 (locus ILSTS-087) to 0.9285 (locus ILSTS-034), with an average value of 0.5485 indicates substantial and very good number of heterozygotes, in the population. The highest PIC value (1.1886) was observed at ILSTS-044 locus and least (0.0768) at ILSTS-065 locus for Marwari goat.

Conclusion: Microsatellite analysis revealed a high level of polymorphism across studied microsatellite markers and informativeness of the markers for genetic diversity analysis studies in Marwari goats. This high level of polymorphism can be utilized to plan future biodiversity studies to exploit the uniqueness and adaptability of this breed to Western Rajasthan. Most studied microsatellite markers proving to be good candidates for genetic characterization and diversity analysis of this breed of goat.

Monday, 20 July 2015

5. Animal health care seeking behavior of pets or livestock owners and knowledge and awareness on zoonoses in a university community - Emmanuel J. Awosanya and H. O. Akande

Veterinary World, 8(7): 841-847

doi: 10.14202/vetworld.2015.841-847

Abstract

Aim: We investigated the attitude of pets or livestock owning households in a university community to animal health care services and assessed the knowledge and awareness level of the residents on zoonoses.

Materials and Methods: Structured questionnaire was used to obtain information on demography, pet or livestock ownership, animal health care seeking behavior, awareness and knowledge of zoonoses from 246 households. We did descriptive statistics and bivariate analysis to determine the level of association in discrete variables between owners and non-owners of pets or livestock at a significant level of p<0.05.

Results: Of the 246 respondents, 80 (32.5%) were either pet or livestock owners. The animal health care seeking behavior of the 80 pets or livestock owners in terms of treatment and vaccination was 70%. Of the 56 (70%) who provided health care services for their animals, about 48 (85.7%) engaged the services of a veterinarian. Dog owning households (42) had the highest frequency of treating their pets against endoparasites (97.6%); ectoparasites (81%) and vaccination against diseases (73.8%). Of the 246 respondents, only 47 (19.1%) have heard of the term zoonoses. Of the considered zoonoses; their awareness of rabies (79.3%) was the highest, followed by Lassa fever (66.3%), the least was pasteurellosis with 18.7%. Having pets or livestock was significantly associated (p=0.04) with rabies awareness. However, there is no significant difference in the level of awareness of zoonoses; knowledge of zoonoses, knowledge of prevention of zoonoses and knowledge of risk of zoonoses between owners and non-owners of pets or livestock.

Conclusion: The animal health care seeking behavior of households with pets or livestock is good and should be encouraged. Public education should be created for other zoonoses aside from rabies, Lassa fever, and avian influenza.

Aim: The objective of this study was to compare two-dimensional (2D) and three-dimensional (3D) study of the pregnant uterus and antenatal development of the fetus.

Materials and Methods: 2D and 3D ultrasound were performed from day 20 to 120 of gestation, twice in week from day 20 to 60 and once in week from day 60 to 120 of gestation on six goats. The ultrasonographic images were obtained using Toshiba, Nemio-XG (Japan) 3D ultrasound machine.

Results: On the 20th day of gestation, earliest diagnosis of pregnancy was done. First 3D ultrasonographic image of the conceptus, through transabdominal approach, was obtained on day 24. On 39th day, clear pictures of conceptus, amniotic membrane, and umbilicus were seen. On 76th day of gestation, internal organs of fetus viz heart, kidney, liver, urinary bladder, and stomach were seen both in 2D and 3D images. 3D imaging showed better details of uterine structures and internal organs of the fetus.

Conclusions: Comparing 3D images with 2D images, it is concluded that 2D was better in visualizing fluid while 3D images were better to view details of attachment of fetus with endometrium.

Aim: To evaluate different thawing temperatures and duration on the post-thaw semen quality of Indian yaks bulls.

Materials and Methods: Semen ejaculates from four different yak bulls were collected using artificial vagina method and extended with tris extender containing 6.4% glycerol at 35°C, cooled gradually from 35°C to 5°C at 1°C/3 min and equilibrated at 4-5°C for 4 h and frozen in French mini straws using a programmable bio-freezer and finally stored in liquid nitrogen.Thawing of frozen semen straws was carried out using three methods i.e., 35°C for 60 s (thawing method I), 37°C for 30 s (thawing method II) and 75°C for 9 s (thawing method III).The post-thaw semen quality parameters assessed were sperm motility, percent live sperm, hypo-osmotic swelling test (HOST)-reacted sperm, acrosomal changes, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in the extracellular media.

Results: The percent sperm motility, total incidence of acrosomal changes, and extracellular release of AST varied significantly (p<0.01) between thawing methods but live sperm and HOST-reacted sperm did not vary significantly between thawing methods.The percent sperm motility of frozen yak semen for thawing method III was significantly (p<0.05) higher than that for thawing methods I and II, the difference between thawing methods I and II being non-significant. The critical difference test revealed that the total incidence of acrosomal changes and extracellular release of AST were significantly (p<0.05) lower when thawing was done using methods I and II than in method III.

Conclusion: On the basis of the present experiment, we can conclude that barring the post-thaw sperm motility, thawing of frozen yak semen in water either at 35°C for 60 s or 37°C for 30 s gives better post-thaw semen quality than at 75°C for 09 s.

Aim: In this work, we aimed to develop maps of modeling geographic distribution correlating to environmental suitability for the two species of scorpions of medical importance at São Paulo State and to develop spatial configuration parameters for epidemiological surveillance of these species of venomous animals.

Materials and Methods: In this study, 54 georeferenced points for Tityus serrulatus and 86 points for Tityus bahiensis and eight environmental indicators, were used to generate species distribution models in Maxent (maximum entropy modeling of species geographic distributions) version 3.3.3k using 70% of data for training (n=38 to T. serrulatus and n=60 to T. bahiensis) and 30% to test the models (n=16 for T. serrulatus and n=26 for T. bahiensis). The logistic threshold used to cut models in converting the continuous probability model into a binary model was the “maximum test sensitivity plus specificity,” provided by Maxent, with results of 0.4143 toT. serrulatus and of 0.3401 to T. bahiensis. The models were evaluated by the area under the curve (AUC), using the omission error and the binomial probability. With the data generated by Maxent, distribution maps were produced using the “ESRI® ArcGIS 10.2.2 for Desktop” software.

Results: The models had high predictive success (AUC=0.7698±0.0533, omission error=0.2467 and p<0.001 for T. serrulatus and AUC=0.8205±0.0390, omission error=0.1917 and p<0.001 for T. bahiensis) and the resultant maps showed a high environmental suitability in the north, central, and southeast of the state, confirming the increasing spread of these species. The environmental variables that mostly contributed to the scorpions species distribution model were rain precipitation (28.9%) and tree cover (28.2%) for the T. serrulatus and temperature (45.8%) and thermal amplitude (12.6%) for the T. bahiensis.

Conclusion: The distribution model of these species of medical importance scorpions in São Paulo State revealed a higher environmental suitability of these species in the regions north, central, and southeast of the state, warning to emergencies actions for prevention and surveillance from scorpion stings in several counties. There is also a need to best conservation strategies related to neighboring territories, with the implementation of new environmental protected areas and measures of spread control of these species in urban areas of several counties.

Aim: The objective of this study was to obtain and compare two-dimensional (2D) and three-dimensional (3D) ultrasonographic images of the urinary bladder in different disease conditions.

Materials and Methods: The present study was conducting on total 10clinical cases of the urinary bladder in dogs. The ultrasound (US) machine used for this study was 3D US machine (Nemio-XG: Toshiba, Japan) having a four-dimensional volumetric probe.

Results: In the present study, the inflamed thickened wall was clearly visible with the distinction of different layers of the urinary bladder wall in some of the cases of cystitis using 2D ultrasonography. In 3D sonogram, the urinary bladder was visualized as a large anechoic structure with no distinction of different layers of the bladder wall. The cystoliths were clearly visible as hyperechoic structures with distal acoustic shadow in 2D sonogram and appeared as a bright echogenic area in 3D sonogram. In case of urinary bladder neoplasia in 2D ultrasonogram, the bladder lumen was found to be occluded with a large growth imaged as focal anechoic areas in the tissue of mixed echogenicity with small hyperechoic dots in this tissue parenchyma. In 3D ultrasonogram, a tissue of mixed echogenicity of pus was also observed.

Conclusion: From the present study it was concluded that 2D and 3D ultrasonography is very helpful for diagnosis of different clinical conditions of the urinary bladder such as cystitis, cystoliths, and urinary bladder neoplasia. The cavity of urinary bladder was more clearly visualized in 3D ultrasonography, but the distinction of different layers of the bladder wall was visualized only in 2D ultrasonography. The distinct shadow of pus and cystoliths were visible in 2D ultrasonogram. The visualization of pus in 3D ultrasonography was done for the first time in present study.